Complex recombination with deletion in the F8 and duplication in the TMLHE mediated by int22h copies during early embryogenesis

Summary

Haemophilia A (HA) is a common X-linked recessive bleeding disorder and almost one half of patients with severe HA are caused by intron 22 inversion (Inv22) in the F8. Inv22 is considered to be almost exclusively of meiotic origin in germ cells during spermatogenesis and only one mosaic Inv22 female carrier with the mutation possibly occurring during mitosis of the embryo has been reported so far. Previously we have identified a novel complex recombination mediated by int22h copies in a sporadic severe HA pedigree and herein we have localised the sequences flanking the breakpoint region using genome walking technique, AccuCopy technique, gene chip and real-time PCR. The disease causing genetic variant registered an 18.1 kb deletion including part of int22h-1 through the intron 23 of F8 and a 113.3 kb duplication of part of int22h-2 through the intron 1 of TMLHE inserted in the religated region of the F8. Two intrinsically linked mechanisms of recombination-dependent DNA replication: microhomology-mediated break-induced replication (MMBIR) followed by break-induced replication (BIR) might be responsible for the incident of the complex recombination during early embryogenesis of the proband’s mother.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Haemophilia A - intron 22 inversion - somatic and germline mosaicism - genomic recombination

• References

• 1 Lenting PJ, van Mourik JA, Mertens K. The life cycle of coagulation factor VIII in view of its structure and function. Blood 1998; 92: 3983-3996.
• 2 Lakich D, Kazazian Jr HH, Antonarakis SE. et al. Inversions disrupting the factor VIII gene are a common cause of severe haemophilia A. Nat Genet 1993; 05: 236-241.
• 3 Rossetti LC, Radic C, Abelleyro MM. et al. Eighteen years of molecular genotyping the hemophilia inversion hotspot: from southern blot to inverse shifting-PCR. Int J Mol Sci 2011; 12: 7271-7285.
• 4 El-Hattab A, Fang W, Jin W. et al. Int22h-1/int22h-2-mediated Xq28 rearrangements: intellectual disability associated with duplications and in utero male lethality with deletions. J Med Genet 2011; 48: 840-850.
• 5 Casana P, Mayo S, Monfort S. et al. Large deletion in the Factor VIII gene (F8) involving segmental duplications in int22h shows no haematological phenotype in female carriers, but may be embryonic lethal in males. Br J Haematol 2012; 158: 138-140.
• 6 Andersen EF, Baldwin EE, Ellingwood S. et al. Xq28 duplication overlapping the int22h-1/int22h-2 region and including RAB39B and CLIC2 in a family with intellectual and developmental disability. Am J Med Genet A 2014; 164A: 1795-1801.
• 7 Jenkins PV, Collins PW, Goldman E. et al. Analysis of intron 22 inversions of the factor VIII gene in severe hemophilia A: implications for genetic counseling. Blood 1994; 84: 2197-2201.
• 8 Ljung RC. Intron 22 inversions and haemophilia. Lancet 1994; 343: 791.
• 9 Rossiter JP, Young M, Kimberland ML. et al. Factor VIII gene inversions causing severe hemophilia A originate almost exclusively in male germ cells. Hum Mol Genet 1994; 03: 1035-1039.
• 10 Theophilus BD, Enayat MS, Green PM. et al. Unique heterozygous intron 22 inversion band pattern in a haemophilic male detected by long polymerase chain reaction (PCR). Thromb Haemost 2008; 99: 774-775.
• 11 Zimmermann MA, Oldenburg J, Muller CR. et al. Unusual genomic rearrangements in introns 1 and 22 of the F8 gene. Hamostaseologie 2011; 31 (01) S69-73.
• 12 Fujita J, Miyawaki Y, Suzuki A. et al. A possible mechanism for Inv22-related F8 large deletions in severe hemophilia A patients with high responding factor VIII inhibitors. J Thromb Haemost 2012; 10: 2099-2107.
• 13 Lannoy N, Grisart B, Eeckhoudt S. et al. Intron 22 homologous regions are implicated in exons 1–22 duplications of the F8 gene. Eur J Hum Genet 2013; 21: 970-976.
• 14 Muhle C, Zenker M, Chuzhanova N. et al. Recurrent inversion with concomitant deletion and insertion events in the coagulation factor VIII gene suggests a new mechanism for X-chromosomal rearrangements causing hemophilia A. Hum Mutat 2007; 28: 1045.
• 15 Kasper CK, Lin JC. Prevalence of sporadic and familial haemophilia. Haemophilia 2007; 13: 90-92.
• 16 Oldenburg J, Rost S, El-Maarri O. et al. De novo factor VIII gene intron 22 inversion in a female carrier presents as a somatic mosaicism. Blood 2000; 96: 2905-2906.
• 17 Levinson B, Lehesjoki AE, de la Chapelle A. et al. Molecular analysis of hemophilia A mutations in the Finnish population. Am J Hum Genet 1990; 46: 53-62.
• 18 Schwaab R, Oldenburg J, Tuddenham EG. et al. Mutations in haemophilia A. Br J Haematol 1993; 83: 450-458.
• 19 Martensson A, Ivarsson S, Letelier A. et al. Origin of mutation in sporadic cases of severe haemophilia A in Sweden. Clin Genet 2016; 90: 63-68.
• 20 Higuchi M, Kochhan L, Olek K. A somatic mosaic for haemophilia A detected at the DNA level. Mol Biol Med 1988; 05: 23-27.
• 21 Leuer M, Oldenburg J, Lavergne JM. et al. Somatic mosaicism in hemophilia A: a fairly common event. Am J Hum Genet 2001; 69: 75-87.
• 22 Brocker-Vriends AH, Briet E, Dreesen JC. et al. Somatic origin of inherited haemophilia A. Hum Genet 1990; 85: 288-292.
• 23 Ding Q, Wu X, Lu Y. et al. AccuCopy quantification combined with pre-amplification of long-distance PCR for fast analysis of intron 22 inversion in haemophilia A. Clin Chim Acta 2016; 458: 78-83.
• 24 Bagnall RD, Giannelli F, Green PM. Int22h-related inversions causing hemophilia A: a novel insight into their origin and a new more discriminant PCR test for their detection. J Thromb Haemost 2006; 04: 591-598.
• 25 You GL, Ding QL, Lu YL. et al. Characterization of large deletions in the F8 gene using multiple competitive amplification and the genome walking technique. J Thromb Haemost 2013; 11: 1103-1110.
• 26 Guo H, Xiong J. A specific and versatile genome walking technique. Gene 2006; 381: 18-23.
• 27 El-Maarri O, Herbiniaux U, Graw J. et al. Analysis of mRNA in hemophilia A patients with undetectable mutations reveals normal splicing in the factor VIII gene. J Thromb Haemost 2005; 03: 332-339.
• 28 Abelleyro MM, Rossetti LC, Radic C. et al. Are int22h-mediated deletions a common cause of hemophilia?. Ann Hematol 2012; 91: 633-636.
• 29 Isrie M, Froyen G, Devriendt K. et al. Sporadic male patients with intellectual disability: contribution of X-chromosome copy number variants. Eur J Med Genet 2012; 55: 577-585.
• 30 Lannoy N, Ravoet M, Grisart B. et al. Five int22h homologous copies at the Xq28 locus identified in intron22 inversion type 3 of the Factor VIII gene. Thromb Res 2016; 137: 224-227.
• 31 Pezeshkpoor B, Rost S, Oldenburg J. et al. Identification of a third rearrangement at Xq28 that causes severe hemophilia A as a result of homologous recombination between inverted repeats. J Thromb Haemost 2012; 10: 1600-1608.
• 32 Lannoy N, Bandelier C, Grisart B. et al. Tandem inversion duplication within F8 Intron 1 associated with mild haemophilia A. Haemophilia 2015; 21: 516-522.
• 33 Zimmermann MA, Oldenburg J, Muller CR. et al. Characterization of duplication breakpoints in the factor VIII gene. J Thromb Haemost 2010; 08: 2696-2704.
• 34 Kasper CK, Buzin CH. Mosaics and haemophilia. Haemophilia 2009; 15: 1181-1186.
• 35 Wylie C. Germ cells. Cell 1999; 96: 165-174.
• 36 Lee JA, Carvalho CM, Lupski JR. A DNA replication mechanism for generating nonrecurrent rearrangements associated with genomic disorders. Cell 2007; 131: 1235-1247.
• 37 Hastings PJ, Ira G, Lupski JR. A microhomology-mediated break-induced replication model for the origin of human copy number variation. PLoS Genet 2009; 05: e1000327.
• 38 Chen JM, Cooper DN, Ferec C. et al. Genomic rearrangements in inherited disease and cancer. Semin Cancer Biol 2010; 20: 222-233.
• 39 Chauvin A, Chen JM, Quemener S. et al. Elucidation of the complex structure and origin of the human trypsinogen locus triplication. Hum Mol Genet 2009; 18: 3605-3614.
• 40 Monfregola J, Napolitano G, Conte I. et al. Functional characterization of the TMLH gene: promoter analysis, in situ hybridization, identification and mapping of alternative splicing variants. Gene 2007; 395: 86-97.
• 41 Celestino-Soper PB, Shaw CA, Sanders SJ. et al. Use of array CGH to detect exonic copy number variants throughout the genome in autism families detects a novel deletion in TMLHE. Hum Mol Genet 2011; 20: 4360-4370.